The proposed studies are designed to develop a cell-free system comprised of nuclei and cytosol isolated from bovine lens epithelial cells. The cell-free system will similar to one in which nuclei and cytosol are derived from rat liver. This cell-free system is designed to specifically study RNA processing and transport. The lens presents and advantage when compared to liver in that it produces relatively high levels of lens-specific mRNAs. This study will focus on the 14S mRNA which codes for the AlphaA2 crystallin. This mRNA can be isolated by sucrose gradient centrifugation of poly(A)-containing RNA prepared from the lens. A 3H-labeled cDNA will be synthesized from this mRNA and used as a probe to quantitate the transport of the mRNA, which codes for AlphaA2 crystallin in the cell-free system. The ability of nuclei isolated from lens epithelial cells to process and transport this specific mRNA will be examined. The participation of cytosolic proteins and other factors in the processing and transport of the mRNA coding for AlphaA2 crystallin will also be studied. Development and characterization of a cell-free system derived from lens epithelial cells should provide a useful tool for studying the processing and transport of individual mRNAs. The initial phase of this study will involve the culturing of bovine lens epithelial cells. This will be done to insure that enough nuclei and cytosol can be obtained in order to carry out the proposed studies.